Characterization of murine TWEAK and its receptor (Fn14) by monoclonal antibodies
Section snippets
Materials and methods
Cells. Mouse T lymphoma L5178Y and myeloma P3U1 (P3X63Ag8U.1) were cultured in RPMI 1640 containing 10% FCS, 100 μg/ml streptomycin and penicillin, and 2 mM glutamine (culture medium). L5178Y-derived transfectants, including mTWEAK/L5178Y and mFn14/L5178Y, were also maintained in the culture medium.
Reagents. mIFN-γ, butylated hydroxyanisole (BHA), and benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z-VAD-fmk) were purchased from BD Bioscience (San Diego, CA), Wako Pure Chemicals (Osaka, Japan),
Characterization of mFn14 transfectants
To characterize the functional properties of mouse Fn14 (mFn14), we first generated stable transfectants expressing full-length mFn14 cDNA. We used L5178Y cells for the transfection, since these cells did not bind CD8-mTWEAK as estimated by flow cytometry (Fig. 1A). The resulting mFn14/L5178Y cells exhibited a high CD8-mTWEAK binding (Fig. 1A). Recently, we have reported that CD8-hTWEAK could induce cell death in human Fn14-transfected L5178Y cells [10]. As shown in Fig. 1B, CD8-mTWEAK could
Acknowledgements
This work was supported by grants from the Human Frontier Science Program, the Ministry of Education, Science, Sports and Culture, and the Ministry of Health, Japan.
References (26)
- et al.
TWEAK, a new secreted ligand in the tumor necrosis factor family that weakly induces apoptosis
J. Biol. Chem.
(1997) - et al.
TWEAK induces angiogenesis and proliferation of endothelial cells
J. Biol. Chem.
(1999) - et al.
A novel TNF receptor family member binds TWEAK and is implicated in angiogenesis
Immunity
(2001) - et al.
Pro-inflammatory effect of TWEAK/Fn14 interaction on human umbilical vein endothelial cells
Biochem. Biophys. Res. Commun.
(2002) - et al.
TWEAK is expressed by glial cells, induces astrocyte proliferation and increases EAE severity
J. Neuroimmunol.
(2002) - et al.
The mitogen-inducible Fn14 gene encodes a type I transmembrane protein that modulates fibroblast adhesion and migration
J. Biol. Chem.
(1999) - et al.
The Fn14 immediate-early response gene is induced during liver regeneration and highly expressed in both human and murine hepatocellular carcinomas
Am. J. Pathol.
(2000) - et al.
Do macrophages kill through apoptosis?
Immunol. Today
(1996) - et al.
A simple, non-chromatographic purification procedure for monoclonal antibodies, isolation of monoclonal antibodies against cytochrome P450 isozymes
J. Immunol. Methods
(1987) - et al.
The transmembrane form of tumor necrosis factor is the prime activating ligand of the 80 kDa tumor necrosis factor receptor
Cell
(1995)